Serwer Philip, Hayes Shirley J, Thomas Julie A, Griess Gary A, Hardies Stephen C
Department of Biochemistry, The University of Texas Health Science Center at San Antonio, San Antonio, TX 78229-3900, USA.
Electrophoresis. 2007 Jun;28(12):1896-902. doi: 10.1002/elps.200600672.
dsDNA viruses with long genomes (>200 kb) are expected to be a major source of novel genes. To rapidly characterize the genomes of newly isolated dsDNA bacteriophages, we develop here a procedure for the PFGE of intact long DNA genomes from bacteriophage particles in unfractionated, infected cell lysates of either liquid or gelled cultures. The DNA used for PFGE is suitable for sequencing after extraction with phenol. The PFGE is tuned to the range of expected DNA lengths. This procedure bypasses the isolation of bacteriophage particles and is useful for PFGE analysis of DNA from dissected zones of bacteriophage plaques.
基因组较长(>200 kb)的双链DNA病毒有望成为新基因的主要来源。为了快速鉴定新分离的双链DNA噬菌体的基因组,我们在此开发了一种程序,用于从液体或凝胶培养物的未分级感染细胞裂解物中的噬菌体颗粒中对完整的长DNA基因组进行脉冲场凝胶电泳(PFGE)。用于PFGE的DNA在用苯酚提取后适合测序。PFGE已调整至预期DNA长度范围。该程序绕过了噬菌体颗粒的分离,可用于对噬菌体噬菌斑解剖区域的DNA进行PFGE分析。